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Intrinsic membrane association of the cytoplasmic tail of influenza virus M2 protein and lateral membrane sorting regulated by cholesterol binding and palmitoylation

Bastian Thaa, +3 more
- 01 Aug 2011 - 
- Vol. 437, Iss: 3, pp 389-397
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TLDR
Pre-conditions of this model, i.e. that the CT interacts with membranes, and that acylation and cholesterol binding affect targeting of M2, showed that palmitoylation and cholesterolbinding differentially affect the intrinsic membrane binding of the amphiphilic helix.
Abstract: 
The influenza virus transmembrane protein M2 is a proton channel, but also plays a role in the scission of nascent virus particles from the plasma membrane. An amphiphilic helix in the CT (cytoplasmic tail) of M2 is supposed to insert into the lipid bilayer, thereby inducing curvature. Palmitoylation of the helix and binding to cholesterol via putative CRAC (cholesterol recognition/interactionaminoacidconsensus)motifsarebelieved to target M2 to the edge of rafts, the viral-budding site. In the present study, we tested pre-conditions of this model, i.e. that the CT interacts with membranes, and that acylation and cholesterol binding affect targeting of M2. M2-CT, purified as a glutathione transferase fusion protein, associated with [ 3 H]photocholesterol and with liposomes. Mutation of tyrosine residues in the CRAC motifs prevented [ 3 H]photocholesterol labelling and reduced liposome binding. M2-CT fused to the yellow fluorescent protein localized to the Golgi in transfected cells; membrane targeting was dependent on CRAC and (to a lesser extent) on palmitoylation. Preparation of giant plasma membrane vesicles from cells expressing full-length M2‐GFP (green fluorescent protein) showed that the protein is partly present in the raft domain. Raft targeting required palmitoylation, but not the CRAC motifs. Thus palmitoylation and cholesterol binding differentially affect the intrinsic membrane binding of the amphiphilic helix.

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Intrinsic membrane association of the cytoplasmic tail of
inuenza virus M2 protein and lateral membrane sorting
regulated by cholesterol binding and palmitoylation
Bastian Thaa, Ilya Levental, Andreas Herrmann, Michael Veit
To cite this version:
Bastian Thaa, Ilya Levental, Andreas Herrmann, Michael Veit. Intrinsic membrane association of
the cytoplasmic tail of inuenza virus M2 protein and lateral membrane sorting regulated by choles-
terol binding and palmitoylation. Biochemical Journal, Portland Press, 2011, 437 (3), pp.389-397.
�10.1042/BJ20110706�. �hal-00608382�
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Biochemical Journal Immediate Publication. Published on 19 May 2011 as manuscript BJ20110706
THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20110706
Accepted Manuscript
Licenced copy. Copying is not permitted, except with prior permission and as allowed by law.
© 2011 The Authors Journal compilation © 2011 Portland Press Limited
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Biochemical Journal Immediate Publication. Published on 19 May 2011 as manuscript BJ20110706
THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20110706
Accepted Manuscript
Licenced copy. Copying is not permitted, except with prior permission and as allowed by law.
© 2011 The Authors Journal compilation © 2011 Portland Press Limited

9--8-
-A /C2-
-    D   -  
  -  - E  E   
    &  A!C2 =3   -
8;,:<;G:<
 -  -  H -8I  -8     A. #C2
,@    -     /    -
&  -  2     @ 3
---A6C
@-@&-A*C2:
/&A)C2
,@/--
        A53 !C2 9    /
@-8-2@
/@8--A .C
@,:-A*C2
/5*>@/.@
;<& 5-;9+<
 #.  ;..E5*<     ;(9< A)C2 9 (9  
-  ; * E*!<    &      8  
- A5C2 9  @ /  --F     
-@--8
A # 6C2
99+/@---;# E6"<
&29@
;G=<9+J&--29
3 - /         - 
@&A *C2@-3-
/ @    J   9+ ;.*3#"<      
@-A )C@
2 9      @    
A 5C @    -  &  -3-
@-@3&2
GG=@&
3 ;HI<    #" A/" / C  -  2 3
 / @ -  3 @     A//C2  @
  -  - H> 
I;(=:(<;K>EL
E#
EMEL
E#
E=>N<A/!C@
 &/O A//
/.C2,@-&@-
(=:(>@-F2
P     3    A/#C    
-A/6C@/
29@-
-/
    '   A//C2     
-       '  @   - 
-   A/* /)C2 9   &  /  -  
-
Biochemical Journal Immediate Publication. Published on 19 May 2011 as manuscript BJ20110706
THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20110706
Accepted Manuscript
Licenced copy. Copying is not permitted, except with prior permission and as allowed by law.
© 2011 The Authors Journal compilation © 2011 Portland Press Limited
 - A//C2    @ @      
&  /           - 
A/5C2
,@     &   -2
G  (=:(     -      
1@(=:('
- @ (3#"A!"! C    (=:(A!/C @ 
@@@2-
-  & /3(9- 
-          D   
A ./5C29-(=:(
-----2
, @  8 @      / -  
@  -   -    (=:( 2 9    
 &  -       -
2,@@/
  @- -        8 
@&-(=:(2@8
@/@-
@>(=:(/2
Biochemical Journal Immediate Publication. Published on 19 May 2011 as manuscript BJ20110706
THIS IS NOT THE VERSION OF RECORD - see doi:10.1042/BJ20110706
Accepted Manuscript
Licenced copy. Copying is not permitted, except with prior permission and as allowed by law.
© 2011 The Authors Journal compilation © 2011 Portland Press Limited
Citations
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What does S-palmitoylation do to membrane proteins?

Sanja Blaskovic, +2 more
- 01 Jun 2013 - 
TL;DR: It is proposed that the direct effects of palmitoylation on transmembrane proteins, however, might be limited to four non‐mutually exclusive mechanistic consequences: alterations in the conformation of trans Membrane domains, association with specific membrane domains, controlled interactions with other proteins and controlled interplay with other post‐translational modifications.
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Association of influenza virus proteins with membrane rafts

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- 25 Jul 2011 - 
TL;DR: The experimental evidence for the viral budozone model is summarized with emphasis on the raft-targeting features of HA, NA, and M2 and the functional importance of raft domains for viral protein transport, assembly and budding, environmental stability, and membrane fusion is reviewed.
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Adaptive Lipid Packing and Bioactivity in Membrane Domains

Erdinc Sezgin, +9 more
- 23 Apr 2015 - 
TL;DR: Evidence is presented that both synthetic and natural plasma membranes assume a wide range of lipid packing states with varying levels of molecular order, which may be adapted and specifically tuned by cells during active cellular processes, as shown for stimulated insulin secretion.
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Model membrane platforms to study protein-membrane interactions

Erdinc Sezgin, +1 more
- 02 Aug 2012 - 
TL;DR: This work summarizes the presently most widely used minimal model membrane systems, namely Supported Lipid Bilayers, Giant Unilamellar Vesicles (GUVs) and Giant Plasma Membrane Vesicles and their applications for protein-membrane interactions.
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Cholesterol-binding site of the influenza M2 protein in lipid bilayers from solid-state NMR

Matthew R. Elkins, +7 more
- 05 Dec 2017 - 
TL;DR: The distance- and orientation-restrained cholesterol-binding site structure shows that cholesterol is stabilized by hydrophobic interactions with the TM helix and polar and aromatic interactions with neighboring amphipathic helices, which suggests that cholesterol mediates M2 clustering to the neck of the budding virus to cause the necessary curvature for membrane scission.
References
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Lipid Rafts As a Membrane-Organizing Principle

Daniel Lingwood, +1 more
- 01 Jan 2010 - 
TL;DR: The evidence for how this principle combines the potential for sphingolipid-cholesterol self-assembly with protein specificity to selectively focus membrane bioactivity is reviewed.
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How proteins produce cellular membrane curvature

Joshua Zimmerberg, +1 more
- 01 Jan 2006 - 
TL;DR: In this paper, the authors classify possible curvature-generating mechanisms that are provided by lipids that constitute the membrane bilayer and by proteins that interact with, or are embedded in, the membrane.
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Structure and Mechanism of the M2 Proton Channel of Influenza A Virus

Jason R. Schnell, +1 more
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TL;DR: The structure of the tetrameric M2 channel in complex with rimantadine, determined by NMR is presented and predicted to counter the effect of drug binding by either increasing the hydrophilicity of the pore or weakening helix–helix packing, thus facilitating channel opening.
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The helical hydrophobic moment: a measure of the amphiphilicity of a helix

David Eisenberg, +2 more
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TL;DR: In this article, the authors defined the mean helical hydrophobic moment, which is defined as the sum of the hydrophobicity of the side chains of a helix of N residues, and reported that trans-membrane helices, which seek surfaces between aqueous and non-polar phases, cluster in different regions of such a plot.
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Palmitoylation: policing protein stability and traffic

Maurine E. Linder, +1 more
- 01 Jan 2007 - 
TL;DR: Recent studies that have provided insights into the mechanisms that mediate the functional consequences of palmitate are reviewed.
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Q1. What are the contributions mentioned in the paper "Intrinsic membrane association of the cytoplasmic tail of influenza virus m2 protein and lateral membrane sorting regulated by cholesterol binding and palmitoylation" ?

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